The capacity of the bread wheat (Triticum aestivum) genome to tolerate introgression from related genomes can be exploited for wheat improvement. A resistance to powdery mildew expressed by a derivative of the cross bread wheat cv. Tähti ⨯ T. militinae (Tm) is known to be due to the incorporation of a Tm segment into the long arm of chromosome 4A. Here, a newly developed in silico method termed RICh (rearrangement identification and characterization) has been applied to characterize the introgression. A virtual gene order, assembled using the GenomeZipper approach, was obtained for the native copy of chromosome 4A; it incorporated 570 4A DArTseq markers to produce a zipper comprising 2,132 loci. A comparison between the native and introgressed forms of the 4AL chromosome arm showed that the introgressed region is located at the distal part of the arm. The Tm segment, derived from chromosome 7G, harbors 131 homoeologs out of the 357 genes present on the corresponding region of Chinese Spring 4AL. The estimated number of Tm genes transferred along with the disease resistance gene was 169. Characterizing the introgression's position, gene content and internal gene order should facilitate not only gene isolation, but may also be informative with respect to chromatin structure and behavior studies.

There is much evidence that mate-choice decisions made by humans are affected by social/contextual information. Women seem to rate men portrayed in a relationship as more desirable than the same men when portrayed as single. Laboratory studies have found evidence suggesting that human mate choice, as in other species, is dependent on the mate choice decisions made by same-sex rivals. Even though non-independent mate choice is an established and well-studied area of mate choice, very few field studies have been performed. This project aims to test whether women’s evaluation of potential mates desirability is dependent/non-independent of same-sex rivals giving the potential mates sexual interest. This is the first field study performed in a modern human’s natural habitat aiming to test for non- independent mate choice in humans.

No desirability enhancement effect was found. The possibilities that earlier studies have found an effect that is only present in laboratory environments or have measured effects other than non-independent mate choice are discussed. I find differences in experimental design to be the most likely reason why the present study failed to detect the effect found in previous studies. This field study, the first of its sort, has generated important knowledge for future experimenters, where the most important conclusion is that major limitations in humans ability to register and remember there surrounding should be taken in consideration when designing any field study investigating human mate choice.

Insertions/deletions (indels) in protein sequences are potential powerful evolutionary markers. However, these characters have rarely been explored systematically at deep phylogenetic levels. Previous analyses of simple (2-state) clade defining indels (CDIs) in universal eukaryotic proteins found none to support any major animal clade. We hypothesized that CDIs might still be found in the remaining population of indels, which we term complex indels. Here, we propose a method for analyzing the simplest class of complex indels the “slightly complex indels”, and use these to investigate deep branches in animal phylogeny. Complex indels with two states, called bi-state indels, show similar evolutionary patterns to singleton simple indels and confirms that insertion mutations are more common than deletions. Exploration of CDIs in 2- to 9-state complex indels shows strong support for all examined branches of fungi and Archaeplastida. Surprisingly, we also found CDIs supporting major branches in animals, particular in vertebrates. We then expanded the search to non-bilaterial animals (Porifera, Cnidaria and Ctenophora). The phylogenetic tree reconstructed by CDIs places the Ctenophore Mnemiopsis leidyi as the deepest branch of animals with 6 CDIs support. Trichoplax adhaerens is closely related to the Bilateria. Moreover, the indel phylogeny shows Nematostella vectensis and Hydra magnipapillata are paraphyletic group and position of Cnidarian branches seems to be problematic in the indel phylogeny because of homoplasy. This might be solved if we discover CDIs from animal specific proteins, which emerged after the universal orthologous proteins.Evolutionary Patterns in Slightly Complex Protein Insertions/Deletions (Indels) and Their Application to the Study of Deep Phylogeny in Metazoa

Genetically encoded, site-specific incorporation of unnatural amino acids (UAA)into proteins through selective recoding of an amber stop codon provides apowerful route for expressing synthetic proteins in living cells. Recoding of theamber stop codon is achieved by introducing an amber suppressortRNA/synthetase pair orthogonal to the endogenous tRNA complement intocells. Methanosarcina is a methane producing archaea with the unusualcapability of suppressing the stop codon (specifically the amber codon). Bysuppressing the amber codon Methanosarcina facilitate the incorporation of thenon-canonical amino acid pyrrolysine (pyl). The suppressing mechanismoriginates from a evolutionary unique Pyrrolysyl-tRNA synthetase (PylRS) and itsmatching tRNApyl. The PylRS has been further evolved and modified to allowincorporation of a wide range of UAAs. Amber suppression is today used tocontrol and study protein function in living cells. By making a series of wellcontrolledexperiments with HEK293T cells we aimed to develop this techniqueinto a robust and general tool for mammalian cell biology. Specifically we weretesting the incorporation of the unnatural amino acid bicyclononyne (BCN) by aset of known PylRS mutants. Our results suggest the mutant aaRS PylRS “AF” isthe most robust and efficient synthetase for BCN. We have improved ambersuppression by determining which factors leads to a more efficient method andsimultaneously decreasing the cost of the method.

We study the performance of stochastic local search algorithms for random instances of the K-satisfiability (K-SAT) problem. We present a stochastic local search algorithm, ChainSAT, which moves in the energy landscape of a problem instance by never going upwards in energy. ChainSAT is a focused algorithm in the sense that it focuses on variables occurring in unsatisfied clauses. We show by extensive numerical investigations that ChainSAT and other focused algorithms solve large K-SAT instances almost surely in linear time, up to high clause-to-variable ratios a; for example, for K = 4 we observe linear-time performance well beyond the recently postulated clustering and condensation transitions in the solution space. The performance of ChainSAT is a surprise given that by design the algorithm gets trapped into the first local energy minimum it encounters, yet no such minima are encountered. We also study the geometry of the solution space as accessed by stochastic local search algorithms.

BACKGROUND: Cadmium (Cd), a ubiquitous food contaminant, has been proposed to be an endocrine disruptor by inducing estrogenic responses in vivo. Several in vitro studies suggested that these effects are mediated via estrogen receptors (ERs).

OBJECTIVE: We performed this study to clarify whether Cd-induced effects in vivo are mediated via classical ER signaling through estrogen responsive element (ERE)-regulated genes or if other signaling pathways are involved.

RESULTS: CdCl2 increased the height of the uterine luminal epithelium in a dose-dependent manner without increasing the uterine wet weight, altering the timing of vaginal opening, or affecting the luciferase activity in reproductive or nonreproductive organs. However, we observed changes in the phosphorylation of mouse double minute 2 oncoprotein (Mdm2) and extracellular signal-regulated kinase (Erk1/2) in the liver after CdCl2 exposure. As we expected, EE2 advanced vaginal opening and increased uterine epithelial height, uterine wet weight, and luciferase activity in various tissues.

Extracts of Plumeria obtusa are widely used in ethnomedicine and have been investigated for a variety of biological activities; however, the antimicrobial activity of P. obtusa flowers is poorly characterized. In this study, the antimicrobial activities of different solvents (petroleum ether, ethyl acetate, chloroform, isobutanol and ethanol) extracts from flowers of P. obtusa were investigated by a disc diffusion method against Gram-positive bacteria, Gram-negative bacteria and a fungus. All extracts exhibited growth inhibition of all microorganisms at variable degrees as measured by relative zones of inhibition, however, the petroleum ether extract was ineffective against Klebsiella pneumonia and ethyl acetate and isobutanol extracts were ineffective against Pseudomonas aeruginosa. The most susceptible Gram-positive bacterium was Bacillus subtilis while the most resistant Gram-positive bacterium was Staphylococcus aureus. Erwinia carotovora was the most susceptible Gram-negative bacterium while P. aeruginosa was highly resistant among the Gram-negative bacteria. In this study, for the first time, we investigated the antimicrobial activity of several different solvent extracts from flowers of P. obtusa against a broad spectrum of human-pathogenic microorganisms. These compounds warrant further investigation by isolation and structural elucidation with the aim to find novel and affordable bioactive compounds for the treatment of infectious diseases.

There are many efforts today trying to mimic the properties of single cells in order to design chips that are as efficient as cells. However, cells are nature's nanotechnology engineering at the scale of atoms and molecules. Therefore, it might be better to vision a microchip that utilizes a single cell as experimentation platform. A novel, so-called Lab-in-a-Cell (LIC) concept is described, where advantage is taken of micro/nanotechnological tools to enable precise control of the biochemical cellular environment and possibility to analyze the composition of single cells.

The importance of living a healthy life in an everyday context is promoted in schools and preschools. The discussion often focuses on what food is healthy, and that one should eat enough but not too much. The connection between food and beverages and their role in the body is seldom discussed. Students’ ideas about how the human body functions have been investigated in several studies but few have focused on young children. In this study, we investigate young children’s conceptions related to this topic and how their ideas develop. Seventy-nine pre- and primary school children, aged 4-11, participated in individual focus interviews wherein the children were asked to draw and explain their understanding. Our results confirm several findings observed by other workers. However, in contrast with earlier studies, 10 of seventeen 4-5 year-old children indicated the stomach, and more than half of those children described how food can be utilized in the body to extract energy. Furthermore, the brain was among the most commonly mentioned organs cross all age groups. Interestingly, the level of expertise varied and did not covariate with age. For example, five of eight of the 4 year-old children draw 5-8 organs, while a single 10 year-old child could only mention three. Similarly, two of thirteen 7-year old children provided an almost completely correct description of the digestive tract and its function, while most of the older children expressed a much less developed understanding. The results reflect the wide range of different conceptual ideas that teachers confront in a day-to-day classroom context.

Understanding of the concept ‘life’ and what characterise ‘living things’ is important as a foundation for learning in biology. In a more general view, this understanding can make children develop awareness, respect and responsibility for life as members of a society and in decision making for sustainable development. The present pilot study aim to investigate 5-6 year old pre-school children’s reasoning and representations about living and nonliving things. In cognitive developmental research, the concept of life is well investigated but, questions still remain regarding how children reason around and represent these concepts. Previous research has found that children have difficulties in including plants as living things. Moreover, it is found that young children include e.g. the sun, clouds and rocks as living things. The methods that have been used are often quantitative and use picture-cards with different objects for the children to categorize. In the present pilot study a modified methodology was applied. Children’s drawings of what they consider as living and non-living were collected and picture-cards were used as point of departure for reasoning. In interviews the children were encouraged to explain and express their ideas. The drawings and the cards mainly worked as a meaning making tool for the children. Results from the study will be presented and discussed.

Trees of Bergvik apple (Bergviksäpple) planted between 2001 and 2007 have shown varying harvest. This study investigates why some trees gets low or no harvest while others get significantly higher harvest. The aim is to identify some reasons why fruiting differs between the trees and if possible suggest and/or enlighten the tree owners what can be done to improve fruiting. The study includes 25 trees, mostly located in Hälsingland, Sweden. The tree owners have answered two questionnaires, first in the spring with questions regarding flowering and the second in the autumn, regarding harvest. The answers to these questionnaires form the foundation for this work.

The result of the study reveals differences in harvest. Older trees and/or trees with good vegetative growth have larger harvest while trees with poor growth in some cases didn’t even get flowers. Younger trees with good growth gave good harvest. This concludes that the single most important factor to get good harvest is good vegetative growth.

Tardigrades are small water-dwelling invertebrates that can live almost anywhere in the world. Even though they are well-known our knowledge about them is still scarce. The aim of this study was therefore to explore our current knowledge about tardigrades by: (1) explore their global phylogeny and biogeography based on bioinformatics (2) screen for tardigrades in select locations of northern Sweden and compare with other Swedish locations, and (3) identify at least one tardigrade from northern Sweden and explore the published biomarkers for further identification. The bulk of this thesis was based on evaluation of the Silva database for analyzing SSU (small subunit) and LSU (large subunit) tardigrade sequences and create phylogenetic trees. Some initial lab work was performed using samples of moss and lichen from Piteå, Vindeln and Öland. Results show that only few countries have been explored with regard to tardigrades, and in Sweden more research have been performed in the south compared to the north. The phylogenetic trees give a rough overview of tardigrade relatedness but many of the sequences need to be improved and more sequence work from additional environments is needed. In the lab tardigrades were only found from the Piteå samples, and one of those was identified as Macrobiotus hufelandi, for which a new biomarker was created. Overall, tardigrade research need to continue and expand to other regions in order to understand how these organisms differ between different environments, and more work is needed to ensure higher quality of sequences added to databases.

The aim of this study was to validate the two presumptive blood tests LMG, LCV and the three visualising blood methods Bluestar Forensics, Lumiscene and the Ruhoff method. The methods’ sensitivity, durability, matrices effects, false positive results and the methods effect on subsequent DNA analysis were studied. DNA analyses were also performed to assess the detection limit of the forensic DNA analysis. Drops of diluted blood were applied on different absorptive matrices and the sensitivity was investigated. The solutions were also placed under different conditions to investigate the durability of the solutions. The solutions were applied upon panels using different chemicals and materials and the false positive results were studied. The DNA analyses were performed by diluting the blood with Bluestar Forensics, the hydrogen peroxide method, the Ruhoff method and deionised water. The study showed that the LMG with a 3 % H2O2 concentration performs the best and it is suited for practical casework. The positive results of LMG was easier to interpret than those of LCV, this is probably due to the fixative agent of the used LCV solution. Bluestar Forensics and Lumiscene did perform similar on the different matrices tested, but the Lumiscene solution had a slightly higher durability. The results strongly indicate that the Ruhoff method can be used without luminol, hence only as a hydrogen peroxide solution (the hydrogen peroxide method). All three visualising blood methods decreases chances of retrieving a positive DNA profile, however the visualising blood methods could be used if the blood cannot be found in any other way. A DNA profile was obtained from the one blood sample analysed at dilution of 1:256 in deionized water.

Background: Maximal oxygen uptake (VO2max) is a determinant of an individual’s ability to handle oxygen during maximal exercise. To measure VO2max expensive equipment and expertise personnel are required. To make the process of measuring VO2max easier, several submaximal and maximal tests have been created in which an estimation of VO2max could be made. Cooper’s 12-minute run (12MR) was created in 1968 and was tested on 115 military men with a mean age of 22 years. Since then the 12MR test has been re- tested and validated towards maximal treadmill tests on several occasions. When an age of 30 years is reached, VO2max starts to decline with 9-10 percent per decade but can be halted by different forms of exercise. With exercise of moderate to high intensity the decline can be halted by almost 50 percent. Objective: To investigate the reliability of estimated VO2max in a test retest scenario of Cooper ́s 12MR on a mixed healthy population between the ages of 28-60. Method: Nine women and five men, healthy subjects with a mean age of 43 ± 8 participated in the present study. A test retest of Cooper ́s 12MR took place with a minimum of seven days between tests. The subjects had to run as many laps as possible on the track during a 12-minute period. Finished laps were then counted and the fraction of the last lap was measured with a measuring wheel and then added to the total distance. To estimate the subjects’ VO2max Cooper ́s table was used. Results: The single measure Intraclass correlation (ICC) that was found, between the estimated VO2max made from the initial test to the retest on Cooper ́s 12MR, was 0.979. ICC showed a small error variance correlation between the tests and was close to the optimal correlation of 1.0. Conclusion: A standardized protocol for performing Cooper ́s 12MR showed good repeatability for estimating VO2max in two separate tests for a mixed population between 28 to 60 years of age.

Microfluidics promises to re-shape the current health-care system by transferring diagnostic tools from central laboratories to close vicinity of the patient (point-of-care). One of the most important operational steps in any diagnostic platform is sample preparation, which is the main subject in this thesis. The goal of sample preparation is to isolate targets of interest from their surroundings. The work in this thesis is based on three ways to isolate bacteria: immune-based isolation, selective cell lysis, size-based separation.

The first sample-preparation approach uses antibodies against lipopolysaccharides (LPS), which are surface molecules found on all gram-negative bacteria. There are two characteristics that make this surface molecule interesting. First, it is highly abundant: one bacterium has approximately a million LPS molecules on its cell-wall. Second, the molecule has a conserved region within all gram-negative bacteria, so using one affinity molecule to isolate disease-causing gram-negative bacteria is an attractive option, particularly from the point of view of sample preparation. The main challenge, however, is antigen accessibility. To address this, we have developed a treatment protocol that improves the capturing efficiency.

The strategy behind selective cell lysis takes advantage of the differences between the blood-cell membrane and the bacterial cell-wall. These fundamental differences make it possible to lyse (destroy) blood-cells selectively while keeping the target of interest, here the bacteria, intact and, what is more important alive. Viability plays an important role in determining antibiotic susceptibility.

Difference in size is another well-used characteristic for sample- separation. Inertial microfluidics can focus size-dependent particle at high flow-rates. Thus, particles of 10 µm diameter were positioned in precise streamlines within a curved channel. The focused particles can then be collected at defined outlets. This approach was then used to isolate white blood cells, which account for approximately 1% of the whole blood. In such a device particles of 2µm diameter (size of bacteria) would not be focused and thereby present at every outlet. To separate bacteria from blood elasto-inertial microfluidics was used. Here, e blood components are diverted to center of the channels while smaller bacteria remain in the side streams and can subsequently be separated.

Hibernation is an extreme physiological challenge for the brown bear (Ursus arctos) in which metabolism is based mainly on lipids. The study objective was to compare plasma lipids in hibernating and active free-ranging brown bears and relate them to arterial histopathology. Blood was drawn from seven immobilized free-ranging brown bears (three females, 23 years old) during hibernation in February and from the same bears while active in June and analyzed by enzymatic and automated hematology methods within 48 hours of sampling. Left anterior descending coronary arteries and aortic arches from 12 bears (six females, 1.512 years old) killed in hunting were examined by histopathology. Total plasma cholesterol decreased from hibernation to the active period (11.08 +/- 1.04 mmol/L vs. 7.89 +/- 1.96 mmol/L, P= 0.0028) as did triglyceride (3.16 +/- 0.62 mmol/L vs. 1.44 +/- 0.27 mmol/L, P= 0.00012) and LDL cholesterol (4.30 +/- 0.71 mmol/L vs. 2.02 +/- 1.03 mmol/L, P= 0.0075), whereas HDL cholesterol was unchanged. No atherosclerosis, fatty streaks, foam cell infiltration, or inflammation were seen in any arterial samples. Brown bears tolerate elevated cholesterol levels, obesity, physical inactivity, and circulatory slow flow during hibernation without signs of -atherosclerosis. This species might serve as a reverse translational model for atherosclerosis resistance.

γ-Aminobutyric acid (GABA) is the main inhibitory neurotransmitter in the brain where it regulates activity of neuronal networks. The receptor for glucagon-like peptide-1 (GLP-1) is expressed in the hippocampus, which is the center for memory and learning. In this study we examined effects of liraglutide, a GLP-1 analog, on GABA signaling in CA3 hippocampal pyramidal neurons.

Methods

We used patch-clamp electrophysiology to record synaptic and tonic GABA-activated currents in CA3 pyramidal neurons in rat hippocampal brain slices.

Results

We examined the effects of liraglutide on the neurons at concentrations ranging from one nM to one μM. Significant changes of the spontaneous inhibitory postsynaptic currents (sIPSCs) were only recorded with 100 nM liraglutide and then in just ≈50% of the neurons tested at this concentration. In neurons affected by liraglutide both the sIPSC frequency and the most probable amplitudes increased. When the action potential firing was inhibited by tetrodotoxin (TTX) the frequency and amplitude of IPSCs in TTX and in TTX plus 100 nM liraglutide were similar.

Conclusions

The results demonstrate that liraglutide regulation of GABA signaling of CA3 pyramidal neurons is predominantly presynaptic and more limited than has been observed for GLP-1 and exendin-4 in hippocampal neurons.

Children are constantly exposed to many chemicals via the products they come in contactwith in their everyday life. One chemical group is phthalates, the most commonly usedplasticisers in the world. Phthalates are used mainly in PVC plastic products like floors, toys,food containers and wallpaper but they are also used in rubber, glue, paint, cables etc. Sincethese chemicals are weakly chemically bound to the PVC they can leak and migrate to the air,food, water and skin. Children are exposed to phthalates mainly through food, but because ofthe hand to mouth behaviour they are also exposed via dust inhalation and dust ingestion.About ten years ago regulations of the most toxic phthalates in toys and child care productswere implemented in the EU and from February 2015 it is a general prohibition for the use ofDEHP (diethylhexyl phthalate), DnBP (di-n-butyl phthalate), DiBP (diisobutyl phthalate) andBBzP (butyl benzyl phthalate). DiNP (diisononyl phthalate) is a relatively new phthalate andit has replaced DEHP to some extent in floors and toys. In phthalate free products DINCH(1,2-cyclohexane dicarboxylic acid diisononyl ester) is one of the plasticizer replacingphthalates. The banned phthalates have been shown to cause adverse effects on reproductionand recent research also investigates links between phthalate exposure and asthma and allergysymptoms in children. More experimental animal studies are needed to further investigate theasthma and allergy correlation.

In the present project dust samples were collected from 30 preschool indoor environments inthe Stockholm city area to evaluate the levels of the six substances mentioned and thephthalate DEP (diethyl phthalate). The main aim of this thesis project was to search forrelations and links between the phthalate concentrations in dust and parameters from theindoor environment in preschools. Studied parameters are e.g. construction year, floor type,cleaning routines and quantity of toys and furniture made of plastic or foam. The project wascommissioned by the Swedish EPA and performed at the Institute of Environmental Medicine(IMM) at Karolinska Institutet.

A negative relation was found between dust phthalate (DEHP and DnBP) concentrationand construction year. Also rooms with old PVC floors had higher concentrations of DEHPand DnBP in dust than rooms with new PVC floors. There was also a trend that dust frompreschools with PVC floors had higher DiNP concentration than dust from preschools withother floor types. The preschools that used foam mattresses for resting had higher DiNPconcentrations than those with no foam mattresses. Most preschools had new foammattresses, which could indicate a more common use of DiNP in new mattresses or mattress2covers compared to old mattresses that contains more DEHP. The four Waldorf preschoolsthat participated had lower DiNP dust concentrations than the other preschools, which wasexpected since Waldorf orientation includes using as little plastic material as possible. Norelation was found between the phthalate dust concentrations and the quantity of toys made ofsoft plastic in the sampled area.

Many preschools had made a plastic inventory where they removed old and soft plastic toysand material. Also many preschools recently replaced old foam mattresses used for resting.This could be due to the big chemical focus in the media and authorities and the brochuresthat have been sent out the last couple of years about what preschool can do to decrease thechemical exposure of children. This interest and awareness seen in the preschools waspositive and hopefully the trend spreads to more preschools. Since children spend a big part oftheir time at preschools it is an important mission for society and the government to decreasethe exposure to hazardous chemicals there. Hopefully what has been done so far is just thebeginning.

The transition to flowering in Arabidopsis is characterized by the sharp and localized upregulation of APETALA1 (AP1) transcription in the newly formed floral primordia. Both the flower meristem-identity gene LEAFY (LFY) and the photoperiod pathway involving the FLOWERING LOCUS T (FT) and FD genes contribute to this upregulation. These pathways have been proposed to act independently but their respective contributions and mode of interaction have remained elusive. To address these questions, we studied the AP1 regulatory region. Combining in vitro and in vivo approaches, we identified which of the three putative LFY binding sites present in the AP1 promoter is essential for its activation by LFY. Interestingly, we found that this site is also important for the correct photoperiodic-dependent upregulation of AP1. In contrast, a previously proposed putative FD-binding site appears dispensable and unable to bind FD and we found no evidence for FD binding to other sites in the AP1 promoter, suggesting that the FT/FD-dependent activation of AP1 might be indirect. Altogether, our data give new insight into the interaction between the FT and LFY pathways in the upregulation of AP1 transcription under long-day conditions.

In a study during the 1970s co-variation of nitrogenase activity and methane formation associated with Sphagnum riparium was observed. This was suggested as evidence for a possible mechanism of hydrogen transfer from cyanobacteria to methanogens. We show experimentally that such a pathway is feasible. In a series of laboratory experiments, using a hydrogenase deficient strain of the heterocystous cyanobacterium Nostoc punctiforme and the hydrogenotrophic methanogen Methanospirillum hungateii in co-cultures, increasing light intensities resulted in elevated nitrogenase activity and methane production. The increase in methane production can be directly deduced from the nitrogenase activity of the N. punctiforme based on hydrogen balance calculations. These experimental results clearly suggest the possible existence of a novel photosynthetically regulated pathway for methane formation.

Female growth, number of eggs and egg growth of Daphnia pulex were studied at four temperatures, 12, 16, 18 and 20°C. I hypothesized that female growth and number of eggs would be lower and the size of the eggs larger in the presence of a predator (kairomones) than in its absence. This effect was expected to be more evidentat lower temperatures. I could not find any effect of kairomones on female growth, number of eggs or egg size, although there was a weak tendency for an effect on number of eggs. There was, however, an effect of temperature, regardless of predator treatment (i. e. presence vs absence of kairomones), on number of eggs, where females produced fewer numbers of eggs with increasing temperature. The effect temperature on the number of eggs was not anticipated. Two of the temperatures, 12 and 18°C, were carried out in darkness due to malfunctioning equipment. In addition, for some unknown reason, a high ratio between the number of ephippia and eggs occurred at at16 and 18°C, regardless of predator treatment. These complicating factors (darkness and ephippia) can have had a negative effect on detecting a response by Daphnia to kairomones.

Metals have been used by humans for thousands of years, and this has resulted in increased concentrations in the biosphere. The environment around point-sources, such as mines and smelters, are of particular concern, as metals may accumulate to high concentrations, and potentially reach levels toxic to the local flora and fauna. This thesis focuses on the effects on pied flycatcher populations of two such point-sources, a lead mine and enrichment plant, and a sulfide ore smelter. Mining activities at the lead mine ceased in 2001 and pied flycatcher populations were assessed before and after the closure. At the sulfide ore smelter, pied flycatchers were studied during the 1980s. Since then, the metal emissions to air from the smelter (e.g. arsenic, cadmium, copper, mercury, lead and zinc) have been greatly reduced (by 93 – 99%).

Pied flycatchers from these two contaminated environments differed in their responses to reduced atmospheric deposition. At the mine site, nestling responses reflected the reduced atmospheric deposition and less lead accumulated in their tissues. However,lead levels were still high enough to cause negative effects on blood status (δ-aminolevulinic acid dehydratase [ALAD], hemoglobin [Hb], hematocrit [ht], and mean cell hemoglobin concentration) and reproduction (reduced clutch size, increased mortality and reduced breeding success), as was observed when the mine was in operation. Along the pollution gradient away from the smelter, nestling concentrations reflected the metal load in the soil pool, accumulating over time, rather than the atmospheric deposition. This resulted in only a minor response to decreased metal deposition (slightly reduced liver lead concentrations at 3.5 – 90 km from the smelter). This suggests that in environments with highly polluted soils, decreased inputs of atmospheric metal deposition have only minor impacts, and recovery from contamination should not be expected within decades.

The high metal concentrations in the vicinity of the smelter contributed to poorer blood status (ALAD, Hb and ht), induced oxidative damage and defenses, and decreased reproduction (increased mortality and reduced breeding success). There were only minor improvements in blood and reproductive variables at 3.5 km from the smelter.

Oxidative stress has been suggested as a mediator in life-history trade-off. By spending more resources on for example reproduction an organism might sacrifice its antioxidant defence. So far, most conclusions on trade-offs between life-history traits and oxidative stress have been drawn from laboratory studies using a few model species and there is a need for studies conducted in natural settings. We investigated associations between markers for antioxidant status (antioxidant enzyme activities and antioxidant levels), body condition, age and reproduction in three species of wild-living passerines. The impact from an anthropogenic stressor (metal pollution) was also assessed. The three bird species showed interspecific variation in their SOD and CAT activities, indicating different pathways to eliminate radicals. The age of females affected both antioxidant status and the breeding performance, indicating the importance of age as a factor in life-history studies. Old birds had lower levels of antioxidants/antioxidant enzyme activities and they produced larger broods/more successful broods, though the latter might be confounded by surviving females having increased fitness. Metal exposure had a negative impact on breeding, and improved breeding outcome was also associated with increased antioxidant defence, but metal exposure was not directly related to the oxidative status of birds, emphasizing that additional stressors might independently affect the same traits. Our results highlight that caution has to be taken when generalizing and extrapolating results to even closely related species. The results support the idea that there is a cost of reproduction, in terms of increased resources spent on antioxidant defence, though this should be confirmed with experimental studies.

Altitudinal gradients offer attractive opportunities for studies of population differentiation in response to environmental heterogeneity. In this thesis, I examined population differentiation along altitudinal gradients by combining common-garden experiments with field studies and experiments in alpine, subalpine and boreal populations of the perennial herb Solidago virgaurea. More specifically, I determined whether leaf physiology in terms of nitrogen concentration and resorption, flowering phenology, flower production and reproductive effort vary along altitudinal gradients.

Nitrogen concentration in green leaves were higher in alpine than in subalpine and boreal populations. These differences persisted when plants were grown from seeds in a common-garden experiment at two sites, suggesting that the differences have a genetic component. There was mixed support for a trade-off between maximized carbon gain through the maintenance of high nitrogen concentration, and minimized nitrogen loss through high resorption.

In their natural habitats alpine populations began flowering later than subalpine populations, but this difference was reversed when plants were grown in a common environment. This suggests that genetic differences among populations counteract environmental effects and reduce phenotypic variation in flowering time among populations. Flowering time thus shows countergradient genetic variation in S. virgaurea.

In a common-garden experiment, boreal populations produced more flowers and had a higher reproductive effort than subalpine and alpine populations indicating habitat-specific genetic differences in reproductive allocation. In a field study, which included three populations, seed set was close to zero in the alpine population, intermediate in the subalpine population, and high in the boreal population. Experimental flower removal showed that seed production was associated with a considerable cost in terms of reduced flowering propensity the following year, but did not support the hypothesis that a large floral display is important for pollination success.

Background: High expression of the polymeric immunoglobulin receptor (PIGR) has previously been associated with a favourable prognosis in a few cancer forms, but its expression and relationship with clinical outcome in epithelial ovarian cancer (EOC) has not yet been reported. The aim of this study was therefore to examine the clinicopathological correlates and prognostic significance of PIGR expression in EOC. Methods: After an initial screening in the Human Protein Atlas portal, a validated antibody was selected for extended analysis of immunohistochemical PIGR expression in tissue microarrays with tumours from 154 incident cases of EOC from two pooled prospective population-based cohorts. Subsets of corresponding benign-appearing fallopian tubes (n = 38) and omental metastases (n = 33) were also analysed. Kaplan-Meier analysis and Cox regression analysis were applied to examine the impact of PIGR expression on overall survival (OS) and ovarian cancer-specific survival (OCSS). Results: PIGR expression was significantly higher in fallopian tubes compared to primary tumours and metastases (p < 0.001) and lower in carcinoma of the serous subtype compared to other carcinomas (p < 0.001). PIGR expression was significantly associated with lower grade (p = 0.001), mucinous histological subtype (p = 0.002), positive progesterone receptor expression (p = 0.009) and negative or low Ki-67 expression (p = 0.003). Kaplan-Meier analysis revealed a significantly improved OS (p = 0.013) and OCSS (p = 0.009) for patients with tumours displaying high expression of PIGR. These associations were confirmed in unadjusted Cox regression analysis (HR = 0.48; 95% CI 0.26-0.87; p = 0.015 for OS and HR = 0.43, 95% CI 0.22-0.82; p = 0.011 for OCSS) but did not remain significant after adjustment for age, grade and clinical stage. Conclusions: This study provides a first demonstration of PIGR expression in human fallopian tubes, primary EOC tumours and metastases. High tumour-specific expression of PIGR was found to be associated with a favourable prognosis in unadjusted, but not in adjusted, analysis. These findings are novel and merit further investigation.